Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli | Thermus thermophilus |
Protein Variants | Comment | Organism |
---|---|---|
A31G/G43A/A709G | optimum temperature is shifted to lower temperatures compared to wild-type enzyme, activity is less temperature dependent than that of the wild-type, resulting in higher activity at temperatures below 60°C, improved turnover-numbers at 40°Cm increase in Km-values, slight loss of thermal stability | Thermus thermophilus |
C275T | similar pH-dependence to that of the wild-type enzyme, improved binding affinities for both D-3-isopropylmalate and NAD+ result in an improved catalytic efficiency, mutant enzyme retains thermal stability | Thermus thermophilus |
G376A | optimum temperature is shifted to lower temperatures compared to wild-type enzyme, activity is less temperature dependent than that of the wild-type, resulting in higher activity at temperatures below 60°C, improved turnover-numbers at 40°C, increase in Km-values, mutant enzyme retains thermal stability | Thermus thermophilus |
G43A | optimum temperature is shifted to lower temperatures compared to wild-type enzyme, activity is less temperature dependent than that of the wild-type, resulting in higher activity at temperatures below 60°C, improved turnover-numbers at 40°C, increase ion Km-values, slight loss of thermal stability | Thermus thermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.001 | - |
isopropylmalate | pH 8.0, 40°C, mutant enzyme C275T | Thermus thermophilus | |
0.0012 | - |
isopropylmalate | pH 8.0, 40°C, mutant enzyme G376A | Thermus thermophilus | |
0.00125 | - |
isopropylmalate | pH 8.0, 40°C, wild-type enzyme | Thermus thermophilus | |
0.0013 | - |
isopropylmalate | pH 8.0, 40°C, mutant enzyme A31G/G43A/A709G | Thermus thermophilus | |
0.0024 | - |
isopropylmalate | pH 8.0, 40°C, mutant enzyme G43A | Thermus thermophilus | |
0.0078 | - |
isopropylmalate | pH 8.0, 70°C, mutant enzyme C275T | Thermus thermophilus | |
0.0089 | - |
isopropylmalate | pH 8.0, 70°C, wild-type enzyme | Thermus thermophilus | |
0.0096 | - |
isopropylmalate | pH 8.0, 70°C, mutant enzyme A31G/G43A/A709G | Thermus thermophilus | |
0.01 | - |
isopropylmalate | pH 8.0, 70°C, mutant enzyme G376A | Thermus thermophilus | |
0.011 | - |
isopropylmalate | pH 8.0, 70°C, mutant enzyme G43A | Thermus thermophilus | |
0.0149 | - |
NAD+ | pH 8.0, 40°C, reaction with 3-D-isopropylmalate, mutant enzyme C275T | Thermus thermophilus | |
0.0258 | - |
NAD+ | pH 8.0, 40°C, isopropylmalate dehydrogenase wild-type | Thermus thermophilus | |
0.157 | - |
NAD+ | pH 8.0, 40°C, mutant enzyme A31G/G43A/A709G | Thermus thermophilus | |
0.16 | - |
NAD+ | pH 8.0, 40°C, mutant enzyme G43A | Thermus thermophilus | |
0.225 | - |
NAD+ | pH 8.0, 40°C, isopropylmalate dehydrogenase cold-adapted mutant T1155 | Thermus thermophilus | |
0.341 | - |
NAD+ | pH 8.0, 70°C, isopropylmalate wild-type | Thermus thermophilus | |
0.82 | - |
NAD+ | at 70°C, mutant enzyme G43A | Thermus thermophilus | |
0.917 | - |
NAD+ | at 70°C, mutant enzyme A31G/G43A/A709G | Thermus thermophilus | |
1.01 | - |
NAD+ | at 40°C, isopropylmalate dehydrogenase cold-adapted mutant P215 | Thermus thermophilus | |
3.56 | - |
NAD+ | 70°C, isopropylmalate dehydrogenase cold-adapted mutant P215 | Thermus thermophilus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermus thermophilus | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2R,3S)-3-isopropylmalate + NAD+ | - |
Thermus thermophilus | 2-oxoisocaproate + NADH + H+ + CO2 | - |
? | |
(2R,3S)-3-isopropylmalate + NAD+ | - |
Thermus thermophilus | (2S)-2-isopropyl-3-oxosuccinate + NADH + H+ | - |
r |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
82 | - |
10 min, 50% inactivation, mutant enzyme A31G/G43A/A709G | Thermus thermophilus |
85 | - |
10 min, 50% inactivation, mutant enzyme G43A | Thermus thermophilus |
87 | - |
10 min, 50% inactivation, wild-type enzyme, mutant enzyme C275T and mutant enzyme G376A | Thermus thermophilus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.36 | - |
NAD+ | pH 8.0, 40°C, wild-type enzyme | Thermus thermophilus | |
2.36 | - |
(2R,3S)-3-isopropylmalate | pH 8.0, 40°C, wild-type enzyme | Thermus thermophilus | |
2.41 | - |
NAD+ | pH 8.0, 40°C, mutant enzyme C275T | Thermus thermophilus | |
2.41 | - |
(2R,3S)-3-isopropylmalate | pH 8.0, 40°C, mutant enzyme C275T | Thermus thermophilus | |
7.2 | - |
NAD+ | pH 8.0, 40°C, mutant enzyme A31G/G43A/A709G | Thermus thermophilus | |
7.2 | - |
(2R,3S)-3-isopropylmalate | pH 8.0, 40°C, mutant enzyme A31G/G43A/A709G | Thermus thermophilus | |
10.7 | - |
NAD+ | pH 8.0, 40°C, mutant enzyme G43A | Thermus thermophilus | |
10.7 | - |
(2R,3S)-3-isopropylmalate | pH 8.0, 40°C, mutant enzyme G43A | Thermus thermophilus | |
17.7 | - |
NAD+ | pH 8.0, 40°C, mutant enzyme G376A | Thermus thermophilus | |
17.7 | - |
(2R,3S)-3-isopropylmalate | pH 8.0, 40°C, mutant enzyme G376A | Thermus thermophilus | |
44.7 | - |
NAD+ | pH 8.0, 70°C, mutant enzyme C275T | Thermus thermophilus | |
44.7 | - |
(2R,3S)-3-isopropylmalate | pH 8.0, 70°C, mutant enzyme C275T | Thermus thermophilus | |
45.8 | - |
NAD+ | pH 8.0, 70°C, wild-type enzyme | Thermus thermophilus | |
45.8 | - |
(2R,3S)-3-isopropylmalate | pH 8.0, 70°C, wild-type enzyme | Thermus thermophilus | |
50.1 | - |
NAD+ | pH 8.0, 70°C, mutant enzyme A31G/G43A/A709G | Thermus thermophilus | |
50.1 | - |
(2R,3S)-3-isopropylmalate | pH 8.0, 70°C, mutant enzyme A31G/G43A/A709G | Thermus thermophilus | |
63.6 | - |
NAD+ | pH 8.0, 70°C, mutant enzyme G43A | Thermus thermophilus | |
63.6 | - |
(2R,3S)-3-isopropylmalate | pH 8.0, 70°C, mutant enzyme G43A | Thermus thermophilus | |
100 | - |
NAD+ | pH 8.0, 70°C, mutant enzyme G376A | Thermus thermophilus | |
100 | - |
(2R,3S)-3-isopropylmalate | pH 8.0, 70°C, mutant enzyme G376A | Thermus thermophilus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Thermus thermophilus |